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Isoamylase (Glycogen 6-glucanohydrolase)

Product code: E-ISAMY

600 Units

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Content: 600 Units
Shipping Temperature: Ambient
Storage Temperature: 2-8oC
Formulation: In 3.2 M ammonium sulphate
Physical Form: Suspension
Stability: > 1 year under recommended storage conditions
Enzyme Activity: Isoamylase
EC Number:
CAZy Family: GH13
CAS Number: 9067-73-6
Synonyms: isoamylase; glycogen 6-alpha-D-glucanohydrolase
Source: Pseudomonas sp.
Molecular Weight: 71,500
Concentration: Supplied at ~ 200 U/mL
Expression: Purified from Pseudomonas sp.
Specificity: Hydrolysis of (1,6)-α-D-glucosidic branch linkages in glycogen, amylopectin and their β-limit dextrins.
Specific Activity: ~ 180 U/mg (40oC, pH 4.0 on oyster glycogen) (equivalent to 16 MU Sigma Units/mg)
Unit Definition: One unit of isoamylase activity is the amount of enzyme required to release one µmole of D-glucose reducing sugar equivalent in the presence of oyster glycogen per min at pH 4.0 and 40oC.
Temperature Optima: 50oC
pH Optima: 4
Application examples: Applications in carbohydrate research and in the food and feeds, and cereals industry.
Method recognition: AOAC Method 2000.11 and GB Standard 5009.245-2016

Pure Isoamylase (Glycogen 6-glucanohydrolase) for use in biochemical enzyme assays and in vitro diagnostic analysis. Isoamylase, Fructanase (E-FRMXPD) and Amyloglucosidase (E-AMGDF) are used in the enzyme hydrolysis step of two validated methods for the determination of polydextrose (a low molar mass dietary fiber) in foods: AOAC method 2000.11 and Chinese GB Standard 5009.245-2016.

Please refer to E-ISAMYHP for high purity enzyme suitable for use in starch structural research.

We have a wide range of other CAZyme products available.

Validation of Methods

Certificate of Analysis
Safety Data Sheet
FAQs Data Sheet

Mechanistic insights into the enhanced texture of potato noodles by incorporation of small granule starches.

Ma, M., Zhang, X., Zhu, Y., Li, Z., Sui, Z. & Corke, H. (2024). International Journal of Biological Macromolecules, 257, 128535.

Potato noodles are a popular food due to their unique texture and taste, but native potato starch often fails to meet consumer demands for precise textural outcomes. The effect of blending small granule (waxy amaranth, non-waxy oat and quinoa) starch with potato starch on the properties of noodles was investigated to enhance quality of noodles. Morphological results demonstrated that small granule starch filled gaps between potato starch granules, some of which gelatinized incompletely. Meanwhile, XRD and FTIR analysis indicated that more ordered structures and hydrogen bonding among starch granules increased with addition of small granule starch. The addition of oat or quinoa starch increased gel elasticity, decreased viscosity of the pastes, and increased the tensile strength of noodles, while addition of 30 % and 45 % waxy amaranth starch did not increase G′ value of gel or tensile strength of noodles. These results indicated that amylose molecules played an important role during retrogradation, and may intertwine and interact with each other to enhance the network structure of starch gel in potato starch blended with oat or quinoa starch. This study provides a natural way to modify potato starch for desirable textural properties of noodle product.

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Development of non-linear prediction model for starch blending.

Park, S. & Kim, Y. R. (2023). LWT, 190, 115567.

Starch blending is a common method for achieving ‘clean label’ with altered properties. However, blending is typically non-additive, making it challenging to predict property changes after mixing. Therefore, this study not only investigated the physicochemical properties of blended starch but also predicted them from individual starches. Six starch samples (rice, corn, potato, tapioca, mungbean, waxy rice) were selected and assessed for individual intrinsic properties (granule size, amylose content, crystallinity, amylopectin chain length). Starch binary blends were then prepared from six starch samples at different blending ratios (100:0, 75:25, 50:50, 25:75, 0:100). Non-additive behaviors were shown in pasting stability, gel storage modulus, and hardness. Notably, the gel storage modulus exceeded expectations, increasing by 140.5% in rice-potato (50:50) and 116.2% in potato-tapioca (50:50) blends. The study also revealed strong correlations among starch properties, suggesting the potential to predict physical properties (e.g., gel hardness, storage modulus) based on intrinsic properties. Developed non-linear models based on the data accurately predicted properties beyond the test set, with a high coefficient of determination (R2 = 0.9). As a result, the models proposed in this study successfully predict physical properties of blended starch, benefiting the food industry interested in clean label starch production and application.

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Digestibility of indica rice and structural changes of rice starch during fermentation by Lactobacillus plantarum.

Tang, Y., Chen, W., Zhu, H., Yi, C., Yuan, J., Liu, Y., Li, Z. & Cheng, H. (2023). LWT, 187, 115392.

Fermentation technology is a useful tool to modifying starch, and the relationship between starch structure and digestibility is a crucial parameter that affects fermentation technology. In this study, the changes in digestibility and starch structure of indica rice during fermentation by Lactobacillus plantarum were investigated. The relationship between digestibility and the changes in starch composition, amylose content, chain length distribution of amylopectin, short-range ordered structure, and relative crystallinity was analyzed. The results showed that during the fermentation of indica rice by L. plantarum, the glycemic index (GI) of the indica rice decreased, and after 60 h of fermentation, the GI of indica rice decreased from 82.88 to 74.53. Additionally, the contents of slowly digestible starch (SDS) and resistant starch (RS) increased. Furthermore, the proportion of short chain of amylopectin increased, which facilitated the intermolecular interaction of short chain to form a compact and orderly structure. This led to an increase in short-range ordered structure and relative crystallinity of indica rice starch. These structural changes promoted the stability of starch structure, resulting in a decrease in rice digestibility. This work has significant implications for the development of low-GI rice and rice-based foods.

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Relationships between starch molecular components and eating and cooking qualities of rice using single-segment substitution lines with different Wx loci.

Lin, L., Wang, Y., Xu, X., Teng, B. & Wei, C. (2023). Journal of Cereal Science, 114, 103765.

Waxy (Wx) gene in rice endosperm synthesizes amylose and amylopectin extra-long chain (ELC). Five rice lines with different Wx locus single-segment substitutions under the same genetic background were investigated for relationships of their starch molecular components and eating and cooking qualities (ECQs) including pasting, thermal, textural and sensory properties. Rice lines with different Wx loci exhibited different ECQs, and their starches had different apparent amylose contents (AACs), true amylose contents (TACs), ELC contents (ELCCs), and amylopectin chain length distributions. The AAC was correlated positively to hardness and cohesiveness of cooked rice, and negatively to adhesiveness and palatability of cooked rice and breakdown viscosity of rice flour. The TAC was correlated positively to gelatinization temperature range and negatively to gelatinization temperature and enthalpy. ELCC showed a positive relationship with pasting peak time of rice flour and chewiness of cooked rice, and could potentially be used as an important reference for rice ECQ evaluation. This study could help the applications of Wx loci in rice quality breeding.

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Structures and digestibility of B-type high-amylose rice starches compared with A-type high-amylose rice starches.

No, J. & Shin, M. (2023). Journal of Cereal Science, 112, 103713.

The starch structures and digestibility of starches prepared from newly developed Korean high amylose rice varieties were investigated. Amylose (AM) contents of Goami, Shingil, Goami 2, and Dodam starches were 25.56, 30.34, 36.33, and 45.78%, respectively. The Goami and Shingil starches were A-type crystallinity, while the Goami 2 and Dodam starches were B-type crystallinity. The molecular weights of AM were lower in the B-type starches (1.64 × 105 and 1.88 × 105) than in the A-type starches (3.88 × 105 and 3.00 × 105). The average chain length (CL) and longer branch CL (DP ≥ 25) of amylopectin (AP) were higher in B-type rice starches than in A-type rice starches. The ratio of 1045/1022 cm-1 and relative crystallinity were lower in the B-type rice starches. These results show that B-type starches have a higher AM content (>35%), gelatinization temperature, resistance to digestibility, and longer branch CL of AP, but a lower AM molecular weight than A-type starches.

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Pho1 cooperates with DPE1 to control short maltooligosaccharide mobilization during starch synthesis initiation in rice endosperm.

Dong, X., Chen, L., Yang, H., Tian, L., Dong, F., Chai, Y. & Qu, L. Q. (2023). Theoretical and Applied Genetics, 136(3), 47.

Plastidial α-glucan phosphorylase is a key factor that cooperates with plastidial disproportionating enzyme to control short maltooligosaccharide mobilization during the initiation process of starch molecule synthesis in developing rice endosperm. Storage starch synthesis is essential for grain filling. However, little is known about how cereal endosperm controls starch synthesis initiation. One of core events for starch synthesis initiation is short maltooligosaccharide (MOS) mobilization consisting of long MOS primer production and excess MOS breakdown. By mutant analyses and biochemical investigations, we present here functional identifications of plastidial α-glucan phosphorylase (Pho1) and disproportionating enzyme (DPE1) during starch synthesis initiation in rice (Oryza sativa) endosperm. Pho1 deficiency impaired MOS mobilization, triggering short MOS accumulation and starch synthesis reduction during early seed development. The mutant seeds differed significantly in MOS level and starch content at 15 days after flowering and exhibited diverse endosperm phenotypes during mid-late seed development: ranging from pseudonormal to shrunken (Shr), severely or excessively Shr. The level of DPE1 was almost normal in the PN seeds but significantly reduced in the Shr seeds. Overexpression of DPE1 in pho1 resulted in plump seeds only. DPE1 deficiency had no obvious effects on MOS mobilization. Knockout of DPE1 in pho1 completely blocked MOS mobilization, resulting in severely and excessively Shr seeds only. These findings show that Pho1 cooperates with DPE1 to control short MOS mobilization during starch synthesis initiation in rice endosperm.

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Comparative Analyses of Grain Quality in Response to High Temperature during the Grain-Filling Stage between Wxa and Wxb under Indica and Japonica Backgrounds.

Fan, X., Sun, X., Yang, R., Chen, S., Li, R., Bian, X., XIA, L. & Zhang, C. (2023). Agronomy, 13(1), 17.

Amylose content controlled by Wx determines rice grain quality, which is easily affected by high temperature. Wxa and Wxb are the two typical Wx alleles in rice, however, their effects on quality formation in response to high temperature under the backgrounds of indica rice and japonica rice have not been systematically compared. In this study, the near-isogenic lines (NILs) of Wxa and Wxb with japonica rice 2661 and indica rice 3611 backgrounds were treated by high temperature during the grain-filling stages. High temperature accelerated the grain ripening process, decreased the thousand-kernel weight, and increased the chalkiness degree of all rice samples. However, these traits of Wx NILs with 3611 background were more susceptible to high temperature than those with 2661 background. Furthermore, high-temperature treatment decreased the amylose contents (AC) and starch viscosities but increased the gelatinization temperature of all the Wx NILs. The 3611-Wxa was atypical Wxa-type rice, whose AC was more sensitive to high temperature. The AC result was consistent with quantitative analysis of GBSSI by Western blot. In addition, the effects of Wx genotype and genetic background on rice physicochemical quality (such as the gel consistencies, starch crystallinity, and the morphological structure of starch grains) in response to high temperature were systematically analyzed. These results have important guiding significance for rice-quality improvement under high-temperature climate.

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Combined Effects of BEIIb and SSIIa Alleles on Amylose Contents, Starch Fine Structures and Physicochemical Properties of Indica Rice.

Hu, Y., Zhang, Y., Yu, S., Deng, G., Dai, G. & Bao, J. (2022). Foods, 12(1), 119.

Starch branching enzyme IIb (BEIIb) and soluble starch synthase IIa (SSIIa) play important roles in starch biosynthesis in cereals. Deficiency in the BEIIb gene produces the amylose extender (ae) mutant rice strain with increased amylose content (AC) and changes in the amylopectin structure. The SSIIa gene is responsible for the genetic control of gelatinization temperature (GT). The combined effects of BEIIb and SSIIa alleles on the AC, fine structures, and physicochemical properties of starches from 12 rice accessions including 10 recombinant inbred lines (RIL) and their two parents were examined in this study. Under the active BEIIb background, starches with the SSIIa-GC allele showed a higher GT than those with the SSIIa-TT allele, resulting from a lower proportion of A chain and a larger proportion of B1 chains in the amylopectin of SSIIa-GC. However, starch with the BEIIb mutant allele (be2b) in combination with any SSIIa genotype displayed more amylose long chains, higher amylose content, B2 and B3 chains, and molecular order, but smaller relative crystallinity and proportion of amylopectin A and B1 chains than those with BEIIb, leading to a higher GT and lower paste viscosities. These results suggest that BEIIb is more important in determining the structural and physicochemical properties than SSIIa. These results provide additional insights into the structure-function relationship in indica rice rather than that in japonica rice and are useful for breeding rice with high amylose content and high resistant starch.

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Insights into the Supramolecular Structure and Degradation Mechanisms of Starch from Different Botanical Sources as Affected by Extrusion-based 3D Printing.

Shahbazi, M., Jäger, H., Ettelaie, R. & Ulbrich, M. (2022). Biomacromolecules, In Press.

Extrusion-based 3D printing has emerged as the most versatile additive manufacturing technique for the printing of practically any material. However, 3D printing of functional materials often activates thermo-mechanical degradation, which affects the 3D shape quality. Herein, we describe the structural changes of eight different starch sources (normal or waxy) as a consequence of the temperature of an extrusion-based 3D printing system through in-depth characterization of their molecular and structural changes. The combination of size-exclusion chromatography, small-angle X-ray scattering, X-ray diffraction, dynamic viscoelasticity measurements, and in vitro digestion has offered an extensive picture of the structural and biological transformations of starch varieties. Depending on the 3D printing conditions, either gelatinization was attained (“moderate” condition) or single-amylose helix formation was induced (“extreme” condition). The stiff amylopectin crystallites in starch granules were more susceptible to thermo-mechanical degradation compared to flexible amorphous amylose. The crystalline morphology of the starch varieties varied from B-type crystallinity for the starch 3D printing at the “moderate” condition to a mixture of C- and V-type crystallinity regarding the “extreme” condition. The “extreme” condition reduced the viscoelasticity of 3D-printed starches but increased the starch digestibility rate/extent. In contrast, the “moderate” condition increased the viscoelastic moduli, decreasing the starch digestion rate/extent. This was more considerable mainly regarding the waxy starch varieties. Finally, normal starch varieties presented a well-defined shape fidelity, being able to form a stable structure, whereas waxy starches exhibited a non-well-defined structure and were not able to maintain their integrity after printing. The results of this research allow us to monitor the degradability of a variety of starch cultivars to create starch-based 3D structures, in which the local structure can be controlled based on the 3D printing parameters.

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Shear and extensional rheological properties of whole grain rye and oat aqueous suspensions.

Malafronte, L., Yilmaz-Turan, S., Dahl, L., Vilaplana, F. & Lopez-Sanchez, P. (2022). Food Hydrocolloids, 137, 108319.

Whole grain flours contain polysaccharides with techno-functional and nutritional properties which make them good candidates as natural texturisers in foods and beverages, thus reducing the use of highly refined ingredients. However, the use of plant components to develop complex fluids and soft materials, requires an enhanced understanding of the relationship between their physicochemical and rheological properties. Here, we systematically investigated the shear and extensional rheological properties of aqueous suspensions of whole grain rye and oat flours. Our results indicated that both types of suspensions (3.5 wt %) showed similar shear thinning behaviour (n = 0.4) however, oat suspensions presented higher viscosity and gel-like behaviour (G'>G'') compared to rye. Additionally, the oat suspensions exhibited an apparent extensional viscosity, which was not present in rye suspensions. The rheological properties of the continuous and disperse phases, separated by centrifugation, were investigated before and after starch hydrolysis and protein removal. Our results indicate that the distinct behaviour of oat suspensions is mainly due to the molecular structure of starch in the liquid phase of i.e oat starch had a higher amylose/amylopectin ratio than rye. Whilst the presence of protein and cell wall polysaccharides in the solid phase contribute to the overall rheology of the suspensions. Furthermore, our results show that the systems do not follow the Cox-Merz rule, indicating that they behaved as suspensions of soft particles rather than macromolecules in solution. Aqueous suspensions of whole grain rye and oat flours showed rheological properties that could be of interest to design low-medium viscosity food and beverage products.

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